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目的：研究钙通道阻滞剂异搏定对环孢素Ａ（ＣＳＡ）所致内皮细胞（ＥＣ）损伤的保护作用。方法：在培养的ＥＣ中加入ＣＳＡ１００μｇ／Ｌ的同时加入异搏定１００μｇ／Ｌ作用２４ｈ，以直接计数粘附细胞数观察细胞分离；光镜、电镜观察细胞形态；Ｈ３－ＴｄＲ掺入法及流式细胞仪观察ＤＮＡ抑制；Ｆｕｒａ－２ＡＭ荧光负载法检测细胞内游离钙（［Ｃａ２＋］ｉ）；放射免疫法检测细胞上清中内皮素（ＥＴ）的含量；斑点杂交检测ＰｒｅｐｒｏＥＴ－１ｍＲＮＡ水平。结果：异搏定可减轻ＣｓＡ对内皮细胞的损伤，加入异搏定的内皮细胞，细胞内游离钙下降及细胞分离减少，超微结构损伤减轻，ＤＮＡ合成抑制改善，异搏定还可明显降低ＣＳＡ介导的前原内皮１ｍＲＮＡ（ＰｒｅｐｒｏＥＴ－１ｍＲＮＡ）水平的升高，使细胞上清中ＥＴ的含量减少。结论：异搏定可有效地改善ＣＳＡ１００μｇ／Ｌ作用２４ｈ所致的ＥＣ细胞毒作用，从而防治其所致的肾毒性和高血压。
AIM: To investigate the protective effect of verapamil, a calcium channel blocker, on endothelial cell injury induced by cyclosporine A (CSA). Methods: CSA100μg / L was added to culture EC with 100μg / L of verapamil for 24h, and the number of adherent cells was counted directly to observe the cell separation. The morphological changes of cells were observed under light microscope and electron microscope. The H3-TdR incorporation and flow cytometry The cells were treated with Fura-2AM to detect the intracellular free calcium ([Ca2 +] i), the content of endothelin (ET) in the cell supernatant was detected by radioimmunoassay, and the level of PreproET-1 mRNA was detected by dot blot hybridization. Results: Verapamil can reduce the damage of CsA to endothelial cells. With the addition of verapamil, the decrease of intracellular free calcium and cell detachment, the decrease of ultrastructure damage and the inhibition of DNA synthesis can significantly reduce verapamil CSA-mediated increase in the level of PreproET-1 mRNA reduces the amount of ET in the cell supernatant. Conclusion: Verapamil can effectively improve EC cytotoxicity induced by CSA 100μg / L for 24 hours, thus preventing and treating nephrotoxicity and hypertension.